Pharmaceutical composition

ABSTRACT

A pharmaceutical composition comprising a tricyclic compound (I) or its pharmaceutically acceptable salt, an oil substance, a surfactant, a hydrophilic substance, water, and optionally a pH control agent, with enhanced stability, absorbability and/or a low irritation potential, is provided.

TECHNICAL FIELD

[0001] This invention relates to a pharmaceutical composition comprisinga tricyclic compound of general formula (I) or a pharmaceuticallyacceptable salt thereof which features stability, excellentabsorbability, and/or a reduced irritation potential. Thispharmaceutical composition is of value for the treatment or preventionof inflammatory or hyperproliferative skin diseases or cutaneousmanifestations of immunologically-mediated diseases.

BACKGROUND ART

[0002] The tricyclic compound (I) and its pharmaceutically acceptablesalt for use in accordance with this invention is known to haveexcellent immunosuppressive activity, antimicrobial activity and otherpharmacological activities and, as such, be of value for the treatmentor prevention of rejection reactions by transplantation of organs ortissues, graft-vs.-host diseases, autoimmune diseases, and infectiousdiseases [Japanese Kokai Tokkyo Koho S61-148181, EP-A-0323042, etc.].

[0003] Particularly, those species of tricyclic compound (I) which aredesignated as FR900506 (=FK506 Substance), FR900520, FR900523, andFR900525 are products produced by microorganisms of the genusStreptomyces, such as Streptomyces tsukubaensis No. 9993 [deposited withNational Institute of Bioscience and Human Technology Agency ofIndustrial Science and Technology (formerly Fermentation ResearchInstitute Agency of Industrial Science and Technology ), at 1-3, Higashi1-chome, Tsukuba-shi, Ibaraki, Japan, date of deposit Oct. 5, 1984,accession number FERM BP-927] or Streptomyces hygroscopicus subsp.yakushimaensis No. 7238 [deposited with National Institute of Bioscienceand Human Technology Agency of Industrial Science and Technology(formerly Fermentation Research Institute Agency of Industrial Scienceand Technology ), at 1-3, Higashi 1-chome, Tsukuba-shi, Ibaraki, Japan,date of deposit Jan. 12, 1985, accession number FERM BP-928]. The FK506Substance of the following chemical formula, in particular, is arepresentative compound.

[0004] Chemical name:17-allyl-1,14-dihydroxy-12-[2-(4-hydroxy-3-methoxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl-11,28-dioxa-4-azatricyclo[22.3.1.0^(4,9)]octacos-18-ene-2,3,10,16-tetraone

[0005] It has been demonstrated that the above-mentioned FK506 Substancehas quite excellent immunosuppressive activity and is useful for thetreatment or prevention of rejection by organ transplantation and thetreatment or prevention of diseases in the field of ophthalmology.

[0006] Japanese Kokai Tokkyo Koho H1-157913 discloses that a solution ofFK506 Substance in ethanol is effective in the suppression ofinflammation and that FK506 Substance can be formulated into a lotion,gel, or cream. However, no specific dosage forms of the kinds aredescribed.

[0007] Japanese Kokai Tokkyo Koho H5-17481 discloses an ointmentcomprising a tricyclic compound (I) or its pharmaceutically acceptablesalt, at least a sufficient amount of a dissolution/absorption promoterto dissolve the same, and an ointment base.

[0008] WO94/28894 discloses a lotion comprising a tricyclic compound (I)or its pharmaceutically acceptable salt, a dissolution/absorptionpromoter, a liquid medium, and optionally an emulsifier and/or athickening (rheology modifier).

[0009] Heretofore, ointments have mainly been used in the treatment ofskin diseases. However, different dosage forms suited to differentclinical manifestations and different application sites are beingdemanded.

[0010] The inventors of this invention have studied possiblepharmaceutical compositions for compounds of general formula (I)inclusive of FK506 Substance and found a dosage form having manydesirable characteristics such as high stability, high transdermalabsorbability, and/or reduced dermal irritancy. Thus, specifically thisinvention is directed to an hydrophilic semi-solid composition forexternal use containing said tricyclic compound.

DISCLOSURE OF INVENTION

[0011] In accordance with this invention there is provided apharmaceutical composition comprising a tricyclic compound (I) or itspharmaceutically acceptable salt, an oil substance, a surfactant, ahydrophilic substance, and water, and further optionally a pH controlagent.

[0012] The tricyclic compound for use in this invention can be expressedby the following general formula (I).

[0013] (wherein each of adjacent pairs of R¹ and R², R³ and R⁴ or R⁵ andR⁶ independently

[0014] (a) is two adjacent hydrogen atoms, or

[0015] (b) may form another bond formed between the carbon atoms towhich they are attached,

[0016] and further, R² may be an alkyl group;

[0017] R⁷ is a hydrogen atom, a hydroxy group, a protected hydroxy groupor an alkoxy group, or an oxo group together with R¹;

[0018] each of R⁸ and R⁹ is independently a hydrogen atom or a hydroxygroup;

[0019] R¹⁰ is a hydrogen atom, an alkyl group, an alkyl groupsubstituted by one or more hydroxy groups, an alkenyl group, an alkenylgroup substituted by one or more hydroxy groups, or an alkyl groupsubstituted by an oxo group;

[0020] X is an oxo group, (a hydrogen atom and a hydroxy group), (ahydrogen atom and a hydrogen atom), or a group represented by theformula —CH₂O—;

[0021] Y is an oxo group, (a hydrogen atom and a hydroxy group) (ahydrogen atom and a hydrogen atom), or a group represented by theformula N—NR¹¹R¹² or N—OR¹³;

[0022] each of R¹¹ and R¹² is independently a hydrogen atom, an alkylgroup, an aryl group or a tosyl group;

[0023] each of R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, R²² and R²³ isindependently a hydrogen atom or an alkyl group;

[0024] each of R²⁰ and R²¹ is independently an oxo group or (R²⁰a and ahydrogen atom) or (R²¹a and a hydrogen atom) in which each of R²⁰a andR²¹a is independently a hydroxy group, an alkoxy group or a grouprepresented by the formula —OCH₂OCH₂CH₂OCH₃, or R²¹a is a protectedhydroxy group, or R²⁰a and R²¹a may together represent an oxygen atom inan epoxide ring;

[0025] n is an integer of 1 or 2; and

[0026] in addition to the above definitions, Y, R¹⁰ and R²³, togetherwith the carbon atoms to which they are attached, may represent asaturated or unsaturated 5- or 6-membered nitrogen, sulfur and/or oxygencontaining heterocyclic ring optionally substituted by one or moregroups selected from the group consisting of an alkyl, a hydroxy, analkyl substituted by one or more hydroxy groups, an alkoxy, a benzyl anda group of the formula —CH₂Se(C₆H₅)).

[0027] The above compound (I) or its pharmaceutically acceptable saltcan be provided by the same technology as that described in the twopatent gazettes referred to above. Particularly, the tricyclic compoundsproduced by fermenting Streptomyces tsukubaensis No. 9993 (FERM BP-927)or Streptomyces hygroscopicus subsp. yakushimaensis No. 7238 (FERMBP-928) are known by the identification nos. of FR-900506, FR-900520,FR-900523, and FR-900525 (Japanese Kokai Tokkyo Koho S61-148181).

[0028] The various definitions given in the above general formula (I),generic and subgeneric examples thereof, and preferred species are nowexplained and shown in detail.

[0029] The term “lower” means, unless otherwise indicated, a grouphaving 1 to 6 carbon atoms.

[0030] Preferable examples of the “alkyl groups” include a straight orbranched chain aliphatic hydrocarbon residue, for example, a lower alkylgroup such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl,neopentyl and hexyl.

[0031] Preferable examples of the “alkenyl groups” include a straight orbranched chain aliphatic hydrocarbon residue having one double-bond, forexample, a lower alkenyl group such as vinyl, propenyl (e.g., allylgroup), butenyl, methylpropenyl, pentenyl and hexenyl.

[0032] Preferable examples of the “aryl groups” include phenyl, tolyl,xylyl, cumenyl, mesityl and naphthyl.

[0033] Preferable protective groups in the “protected hydroxy groups”are 1-(lower alkylthio)(lower)alkyl group such as a loweralkylthiomethyl group (e.g., methylthiomethyl, ethylthiomethyl,propylthiomethyl, isopropylthiomethyl, butylthiomethyl,isobutylthiomethyl, hexylthiomethyl, etc.), more preferably C₁-C₄alkylthiomethyl group, most preferably methylthiomethyl group;

[0034] trisubstituted silyl group such as a tri(lower)alkylsilyl (e.g.,trimethylsilyl, triethylsilyl, tributylsilyl, tert-butyldimethylsilyl,tri-tert-butylsilyl, etc.) or lower alkyl-diarylsilyl (e.g.,methyldiphenylsilyl, ethyldiphenylsilyl, propyldiphenylsilyl,tert-butyldiphenyl-silyl, etc.), more preferably tri(C,-C₄)alkylsilylgroup and C₁-C₄ alkyldiphenylsilyl group, most preferablytert-butyldimethylsilyl group and tert-butyldiphenylsilyl group; and anacyl group such as an aliphatic, aromatic acyl group or an aliphaticacyl group substituted by an aromatic group, which are derived from acarboxylic acid, sulfonic acid or carbamic acid.

[0035] Examples of the aliphatic acyl groups include a lower alkanoylgroup optionally having one or more suitable substituents such ascarboxy, e.g., formyl, acetyl, propionyl, butyryl, isobutyryl, valeryl,isovaleryl, pivaloyl, hexanoyl, carboxyacetyl, carboxypropionyl,carboxybutyryl, arboxyhexanoyl, etc.; acyclo(lower)alkoxy(lower)alkanoyl group optionally having one or moresuitable substituents such as lower alkyl, e.g., cyclopropyloxyacetyl,cyclobutyloxypropionyl, cycloheptyloxybutyryl, menthyloxyacetyl,menthyloxypropionyl, menthyloxybutyryl, menthyloxypentanoyl,menthyloxyhexanoyl, etc.; a camphorsulfonyl group; or a loweralkylcarbamoyl group having one or more suitable substituents such ascarboxy or protected carboxy, for example, carboxy(lower)alkylcarbamoylgroup (e.g., carboxymethylcarbamoyl, carboxyethylcarbamoyl,carboxypropylcarbamoyl, carboxybutylcarbamoyl, carboxypentylcarbamoyl,carboxyhexylcarbamoyl, etc.),tri-lower)alkylsilyl(lower)alkoxycarbonyl(lower)alkylcarbamoyl group(e.g., trimethylsilylmethoxycarbonylethylcarbamoyl,trimethylsilylethoxycarbonylpropylcarbamoyl,triethylsilylethoxycarbonylpropylcarbamoyl,tert-butyldimethylsilylethoxycarbonylpropylcarbamoyl,tri-methylsilylpropoxycarbonylbutylcarbamoyl, etc.) and so on.

[0036] Examples of the aromatic acyl groups include an aroyl groupoptionally having one or more suitable substituents such as nitro, e.g.,benzoyl, toluoyl, xyloyl, naphthoyl, nitrobenzoyl, dinitrobenzoyl,nitronaphthoyl, etc.; and an arenesulfonyl group optionally having oneor more suitable substituents such as halogen, e.g., benzenesulfonyl,toluenesulfonyl, xylenesulfonyl, naphthalenesulfonyl,fluorobenzenesulfonyl, chlorobenzenesulfonyl, bromobenzenesulfonyl,iodobenzenesulfonyl, etc.

[0037] Examples of the aliphatic acyl groups substituted by an aromaticgroup include ar(lower)alkanoyl group optionally having one or moresuitable substituents such as lower alkoxy or trihalo(lower)alkyl, e.g.,phenylacetyl, phenylpropionyl, phenylbutyryl,2-trifluoromethyl-2-methoxy-2-phenylacetyl,2-ethyl-2-trifluoromethyl-2-phenylacetyl,2-trifluoromethyl-2-propoxy-2-phenylacetyl, etc.

[0038] More preferable acyl groups among the aforesaid acyl groups areC₁-C₄ alkanoyl group optionally having carboxy,cyclo(C₅-C₆)alkoxy(C₁-C₄)alkanoyl group having two (C₁-C₄) alkyls at thecycloalkyl moiety, camphorsulfonyl group, carboxy(C₁-C₄)alkylcarbamoylgroup, tri(C₁-C₄)alkylsilyl (C₁-C₄)alkoxycarbonyl-(C₁-C₄)alkylcarbamoylgroup, benzoyl group optionally having one or two nitro groups,benzenesulfonyl group having halogen, or phenyl (C₁-C₄)alkanoyl grouphaving C₁-C₄ alkoxy and trihalo(C₁-C₄)alkyl group. Among these, the mostpreferable ones are acetyl, carboxypropionyl, menthyloxyacetyl,camphorsulfonyl, benzoyl, nitrobenzoyl, dinitrobenzoyl,iodobenzenesulfonyl and 2-trifluoromethyl-2-methoxy-2-phenylacetyl.

[0039] Preferable examples of the “5- or 6-membered nitrogen, sulfurand/or oxygen containing heterocyclic ring” include a pyrrolyl group anda tetrahydrofuryl group.

[0040] The pharmaceutically acceptable salt of the compound (I) includesconventional non-toxic and pharmaceutically acceptable salt such as thesalt with inorganic or organic bases, specifically, an alkali metal saltsuch as sodium salt and potassium salt, an alkali earth metal salt suchas calcium salt and magnesium salt, an ammonium salt and an amine saltsuch as triethylamine salt and N-benzyl-N-methylamine salt.

[0041] With respect to the compound (I), it is to be understood thatthere may be conformers and one or more stereoisomers such as opticaland geometrical isomers due to asymmetric carbon atom(s) or doublebond(s), and such conformers and isomers are also included within thescope of the present invention.

[0042] The compound of the formula (I) or its pharmaceuticallyacceptable salt can be in the form of a solvate, which is includedwithin the scope of the present invention. The solvate preferablyinclude a hydrate and an ethanolate.

[0043] The preferred examples of the tricyclic compound (I) is the one,wherein each of adjacent pairs of R³ and R⁴ or R⁵ and R⁶ independentlyform another bond formed between the carbon atoms to which they areattached;

[0044] each of R⁸ and R²³ is independently a hydrogen atom;

[0045] R⁹ is a hydroxy group;

[0046] R¹⁰ is a methyl group, an ethyl group, a propyl group or an allylgroup;

[0047] X is (a hydrogen atom and a hydrogen atom) or an oxo group;

[0048] Y is an oxo group;

[0049] each of R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, and R²² is a methyl group;

[0050] each of R²⁰ and R²¹ is independently (R²⁰a and a hydrogen atom)or (R²¹a and a hydrogen atom) in which each of R²⁰a and R²¹a is ahydroxy group or an alkoxy group, or R²¹a is a protected hydroxy group;and

[0051] n is an integer of 1 or 2.

[0052] FK506 Substance is the most preferable compound belonging to thetricyclic compound (I). Other preferable compounds are listedhereinbelow.

[0053]1,14-Dihydroxy-12-[2-(4-hydroxy-3-methoxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy-13,17,19,21,27-pentamethyl-11,28-dioxa-4-azatricyclo[22.3.1.0^(4,9)]octacos-18-ene-2,3,10,16-tetraone,

[0054]12-[2-(4-acetoxy-3-methoxycyclohexyl)-1-methylvinyl]-17-allyl-1,14-dihydroxy-23,25-dimethoxy-13,19,21,27-tetramethyl-11,28-dioxa-4-azatricyclo[22.3.1.0^(4,9)]octacos-18-ene-2,3,10,16-tetraone,

[0055]17-allyl-1,14-dihydroxy-23,25-dimethoxy-13,19,21,27-tetramethyl-12-[2-[4-(3,5-dinitrobenzoyloxy)-3-methoxycyclo-hexyl]-1-methylvinyl]-11,28-dioxa-4-azatricyclo[22.3.1.0^(4,9)]-octacos-18-ene-2,3,10,16-tetraone,

[0056]17-allyl-12-[2-[4-[(−)-2-trifluoromethyl-2-methoxy-2-phenylacetoxy]-3-methoxycyclohexyl]-1-methylvinyl]-1,14-dihydroxy-23,25-dimethoxy-13,19,21,27-tetramethyl-11,28-dioxa-4-azatricyclo[22.3.1.0^(4,9)]octacos-18-ene-2,3,10,16-tetraone.

[0057]17-ethyl-1,14-dihydroxy-12-[2-(4-hydroxy-3-methoxycyclo-hexyl)-1-methylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl-11,28-dioxa-4-azatricyclo[22.3.1.0^(4,9)]octacos-18-ene-2,3,10,16-tetraone(FR⁹⁰⁰⁵²⁰), and

[0058]17-ethyl-1,14,20-trihydroxy-12-[2-(3,4-dihydroxycyclo-hexyl)-1-methylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl-11,28-dioxa-4-azatricyclo[22.3.1.04,9]octacos-18-ene-2,3,10,16-tetraone.

[0059] The oil substance for use in this invention need only to be asubstance capable of dissolving tricyclic compound (I) or itspharmaceutically acceptable salt. Preferable ones are, for example,fatty acid esters and alcohols.

[0060] Monohydric Alcohol Fatty Acid Esters

[0061] (isopropyl myristate, ethyl myristate, butyl myristate, isocetylmyristate, octyldodecyl myristate, isopropyl palmitate, isostearylpalmitate, isopropyl isostearate, isocetyl isostearate, butyl stearate,isocetyl stearate, cetyl isooctanoate, ethyl linoleate, isopropyllinoleate, hexyl laurate, ethyl oleate, decyl oleate, oleyl oleate,octyl dodecyl myristate, hexyl decyl dimethyloctanoate, octyl dodecylneodecanoate, etc.)

[0062] Dibasic Acid Diesters

[0063] (diisopropyl adipate, dimethyl adipate, diethyl adipate,diisobutyl adipate, diethyl sebacate, diisopropyl sebacate, dipropylsebacate, diethyl phthalate, diethyl pimelate, etc.)

[0064] Alcohols

[0065] (oleyl alcohol, cetanol, stearyl alcohol, 2-octyldodecanol, etc.)

[0066] In this invention the above-mentioned oil substances can be usedindependently or in a combination of two or more species.

[0067] Particularly from the standpoint of stability of the dosage form,absorbability and/or irritation potential, it is preferable, in manyinstances, to use a plurality of such oil substances in combination. Thepreferred combination may for example be a mixture of a monohydricalcohol fatty acid ester (e.g. isopropyl myristate) and a dibasic aciddiester (e.g. diethyl sebacate) in a suitable weight ratio (e.g.0.1˜10:1 (w/w), preferably 0.5˜2:1 (w/w)). The most preferable weightratio thereof is 1:1 (w/w).

[0068] The proportion of the oil substance in the pharmaceuticalcomposition is preferably 2˜50% (w/w), more preferably 10˜40%, and mostpreferably 20˜30%.

[0069] The surfactant for use in this invention is now described.

[0070] The surfactant that can be used includes pharmaceuticallyacceptable ionic or nonionic surfactants but is preferably a nonionicsurfactant with an HLB number of not less than 10. More preferably, thefollowing ether series and ester series surfactants can be mentioned.

[0071] Ethers

[0072] Polyoxyethylene alkyl ethers

[0073] (polyoxyethylene oleyl ether, polyoxyethylene stearyl ether,polyoxyethylene cetyl ether, polyoxyethylene lauryl ether (LauromacrogolJ.P.), polyoxyethylene behenyl ether, etc.)

[0074] Esters

[0075] Polyoxyethylene sorbitan fatty acid esters

[0076] Polyoxyethylene sorbitan monostearate, polyoxyethylene sorbitanmonooleate, etc.

[0077] (e.g. Tween 20, Tween 40, Tween 60, Tween 65, Tween 80, etc., alltrademarks)

[0078] Polyethylene Glycol Fatty Acid Esters

[0079] (polyethylene glycol monooleate, polyethylene glycol monostearate(e.g. polyoxyl stearate 40 J.P.), polyethylene glycol monolaurate, etc.)

[0080] Pentaglycerol fatty acid esters

[0081] Pentaglycerol monolaurate, pentaglycerol monomyristate,pentaglycerol monooleate, pentaglycerol monostearate, etc.

[0082] Glycerol fatty acid esters

[0083] Glyceryl monostearate etc.

[0084] In working this invention, the above-mentioned surfactants can beused independently or in a combination of two or more species. Theproportion of the surfactant in the pharmaceutical composition ispreferably 0.1˜15% (w/w) and more preferably 0.5˜5%(w/w).

[0085] The hydrophilic substance for use in this invention may be anysubstance that is pharmaceutically acceptable and capable of impartingviscosity to liquids, thus including the following organic or inorganichydrophilic substances.

[0086] (1) Organic substances

[0087] {circle over (1)} Natural polymers . . . gum arabic, guar gum,carrageenin, gum tragacanth, pectin, starch, gum xanthan, gelatin,casein, dextrin, cellulose

[0088] {circle over (2)} Semi-synthetic polymers . . . methylcellulose,ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose,carboxymethylcellulose sodium, carboxymethylcellulose calcium,carboxymethylstarch, sodium alginate, propylene glycol alginate,

[0089] {circle over (3)} Synthetic polymers . . . carboxyvinyl polymer(Carbopol), polyvinyl alcohol, polyvinylpyrrolidone, polyethyleneglycol, polyvinyl methyl ether, sodium polyacrylate

[0090] (2) Inorganic substances

[0091] Bentonite, synthetic magnesium silicate, magnesiumaluminosilicate, silicon oxide, etc.

[0092] The proportion of the hydrophilic substance in the pharmaceuticalcomposition of this invention is selected according to the desiredviscosity of the pharmaceutical composition and is preferably 0.1˜10%(w/w) and more preferably 0.5˜2%.

[0093] Furthermore, in the instant invention, the pharmaceuticalcomposition is preferably maintained at a constant pH from safety pointsof view. Therefore, a pH control agent such as a buffer, an aqueoussolution of sodium hydroxide, or the like can be added in a suitableamount. The pH range is preferably 3.5˜6 and more preferably 4˜5.

[0094] Where necessary, in addition to the above ingredients, theconventional excipient (e.g. petrolatum, propylene glycol, etc.),stabilizer (antioxidant etc.), coloring agent, sweetener, flavor,diluent, antiseptic (e.g. parahydroxybenzoates, benzalkonium chloride,sorbic acid, etc.), and other drug substances effective against diseasesof the skin can be added.

[0095] Meanwhile, the pharmaceutical composition of this invention canbe produced by a process comprising the following steps.

[0096] (1) A step which comprises preparing a solution composed of saidtricyclic compound (I) or pharmaceutically acceptable salt thereof, oilsubstance, and surfactant;

[0097] (2) a step which comprises mixing the solution with water to givean emulsion; and

[0098] (3) a step which comprises mixing the emulsion with saidhydrophilic substance, and further optionally pH control agent, withstirring. As an alternative, this step may comprise mixing saidhydrophilic substance, and further optionally pH control agent, withwater and, then, mixing the premix with the emulsion prepared as abovewith stirring.

[0099] The step (1) is preferably carried out at elevated temperature,for example at 50˜90° C., preferably 60˜80° C.

[0100] In step (2), the water for use is also preferably heated ahead oftime to a temperature close to that of the solution prepared in step (1), and the resulting emulsion is preferably cooled to a suitabletemperature, for example 30° C., before step (3) is carried out.

[0101] The dosage of tricyclic compound (I) or its pharmaceuticallyacceptable salt depends on the individual patient's age and the type andseverity of disease but the usual daily therapeutic dose is about0.001˜1000 mg, preferably about 0.005˜500 mg, and more preferably about0.01˜100 mg, as the active ingredient. Generally, an average of about0.01 mg, 0.05 mg, 0.1 mg, 0.5 mg, 1 mg, 5 mg, 10 mg, 25 mg, or 50 mg perdose is administered.

[0102] The recommended proportion of tricyclic compound (I) in thepharmaceutical composition is 0.001˜20% (w/w), preferably 0.01˜10%(w/w),of the total composition.

EXAMPLES

[0103] The following examples illustrate the present invention infurther detail, it being to be understood that those examples are notintended to limit the scope of the invention.

Example 1

[0104] The following ingredients (a), (b), and (c) were first admixed at60˜80° C. to prepare a solution, to which (d) previously warmed to60˜80° C. was added. The mixture was evenly emulsified with a homomixerand the resulting emulsion was cooled to 30° C. Then, (e) was added tothe emulsion and the whole mixture was stirred to provide apharmaceutical composition in the form of an easily spreadable cream[formulation (1)]. Formulation (1) (% w/w) (a) FK506 Substance 0.1 (b)Isopropyl myristate 25.0 (c) Polyoxyethylene[5.5] 5.0 cetyl ether (d)Purified water 68.9 (e) Carbopol 940 1.0

Example 2

[0105] A pharmaceutical composition (formulation (2)) was preparedaccording to a similar manner to that of Example 1, except that, afteraddition of (e), the mixture was adjusted to pH about 4.0 with asuitable amount of 1N-aqueous solution of NaOH (f).

Example 3

[0106] Pharmaceutical compositions (formulations (3) and (4)) wereprepared according to a similar manner to that of Examples 1 and 2.Formulation (3) Formulation (4) (% w/w) (% w/w) (a) FK506 Substance 0.10.1 (b) Isopropyl myristate 25.0 25.0 (c) Polyoxyethylene [73] 5.0 —cetyl ether (c) Polyoxyethylene [10] — 5.0 oleyl ether (d) Purifiedwater 68.9 68.9 (e) Carbopol 940 1.0 1.0 (f) 1N NaOH aq. sol. q.s. q.s.

Example 4

[0107] Pharmaceutical compositions (formulations (5)-(8)) were preparedaccording to a similar manner to that of Examples 1 and 2. (5) (6) (7)(8) Formulation (% w/w) (% w/w) (% w/w) (% w/w) (a) FK506 Substance 0.10.1 0.1 0.1 (b) Isopropyl 25.0 25.0 25.0 25.0 myristate (c)Pentaglycerol 5.0 — — — monolaurate (c) Pentaglycerol — 5.0 — —monomyristate (c) Pentaglycerol — — 5.0 — monooleate (c) Pentaglycerol —— — 5.0 monostearate (d) Purified water 68.9 68.9 68.9 68.9 (e) Carbopol940 1.0 1.0 1.0 1.0 (f) 1N-NaOH aq. sol. q.s. q.s. q.s. q.s.

Example 5

[0108] Pharmaceutical compositions (formulations (9)-(12)) were preparedaccording to a similar manner to that of Examples 1 and 2. (9) (10) (11)(12) Formulation (% w/w) (% w/w) (% w/w) (% w/w) (a) FK506 Substance 0.10.1 0.1 0.1 (b) Isopropyl 25.0 25.0 25.0 25.0 myristate (c) Glycerin 5.0— — — monostearate (c) Polyethylene glycol — 5.0 — — monostearate (c)Polyoxyethylene- — — 5.0 — [20] sorbitan monostearate (c)Polyoxyethylene- — — — 5.0 [20] sorbitan monooleate (d) Purified water68.9 68.9 68.9 68.9 (e) Carbopol 940 1.0 1.0 1.0 1.0 (f) 1N-NaOH aq.sol. q.s. q.s. q.s. q.s.

Example 6

[0109] Pharmaceutical compositions (formulations (13)-16)) were preparedaccording to a similar manner to that of Examples 1 and 2. (13) (14)(15) (16) Formulation (% w/w) (% w/w) (% w/w) (% w/w) (a) FK506Substance 0.1 0.1 0.1 0.1 (b) Diethyl sebacate 25.0 25.0 — 25.0 (b)Oleyl alcohol — — 25.0 — (c) Polyoxyethylene- 5.0 — — — [10] behenylether (c) Polyoxyethylene- — 5.0 — — [5.5] cetyl ether (c)Polyoxyethylene- — — 5.0 — [21] lauryl ether (c) Polyoxyethylene- — — —5.0 [20] sorbitan monooleate (d) Purified water 68.9 68.9 68.9 68.9 (e)Carbopol 940 1.0 1.0 1.0 1.0 (f) 1N-NaOH aq. sol. q.s. q.s. q.s. q.s.

Example 7

[0110] The following ingredients (a), (b), and (c) were admixed at60˜80° C. to prepare a solution, to which (d₁) preheated to 60˜80° C.was added. The mixture was evenly homogenized with a homomixer and,then, cooled to 30° C.

[0111] To this emulsion was added a gel prepared from (e), (d₂), and (f)and the mixture was stirred thoroughly to provide a pharmaceuticalcomposition in the form of a well-spreadable cream [formulation (17)].Formulation (17) (% w/w) (a) FK506 Substance 0.1 (b) Isopropyl myristate25.0 (c) Polyoxyethylene [5.5] 5.0 cetyl ether (d₁) Purified water 34.4(e) Carbopol 940 1.0 (d₂) Purified water 34.5 (f) 1N-NaOH aq. sol. q.s.

Example 8

[0112] A pharmaceutical composition (formulation (18)) was preparedaccording to a similar manner to that of Example 7. Formulation (18) (%w/w) (a) FK506 Substance 0.1 (b) Diethyl sebacate 25.0 (c)Polyoxyethylene [20] 5.0 sorbitan monooleate (d₁) Purified water 34.4(e) Carbopol 940 1.0 (d₂) Purified water 34.5 (f) 1N-NaOH aq. sol. q.s.

Example 9

[0113] Pharmaceutical compositions (formulations (19)-(21)) wereprepared according to a similar manner to that of Example 7. (19) (20)(21) Formulation (% w/w) (% w/w) (% w/w) (a) FK506 Substance 0.03 0.10.3 (b₁) Isopropyl myristate 20 20 20 (b₂) Diethyl sebacate 20 20 20 (c)polyoxyethylene [20] 2.5 2.5 2.5 sorbitan monooleate (d₁) + (d₂)Purified water 56.47 56.4 56.2 (e) Carbopol 940 1.0 1.0 1.0 (f) 1N-NaOHaq. sol. q.s. q.s. q.s.

Example 10

[0114] The protocol and results of a transdermal absorbabilityexperiment using pharmaceutical compositions of the present inventionare described below.

[0115] Transdermal Absorbability Test

[0116] Using the formulations 2, 3, 7, 12, and 20 prepared in theforegoing examples, an in vivo transdermal absorbability experiment wasperformed.

[0117] As experimental animals, 3 male 7-week-old SD rats were used.With the rat in supine position on a stereotactic apparatus, the haircoat in the abdominal region was clipped off with an electric clipperand, then, a depilatory cream (Eva-Cream, manufactured by Tokyo TanabePharmaceutical) was applied. The skin area thus treated was washed withwater 10 minutes after application of the cream for depilation. The ratwas returned to its cage and allowed to rest for 24 hours. Thereafter,the rat was in supine position on the stereotactic apparatus, arectangular area measuring 2.5 cm×4 cm was marked off at four corners onthe depilated abdominal area of the rat and 50 mg of the test sample wasapplied over the square. At 1, 3, 5, 8, and 24 hours after thisapplication, 0.3 ml of blood was drawn from the subclavian vein with anEDTA-pretreated syringe. After thorough mixing with EDTA, the blood wasstored frozen until assayed.

[0118] Using the above blood samples, the whole blood FK506 Substanceconcentration was determined by enzyme-linked immunosorbent assay usingperoxidase as the enzyme (e.g. shown in Japanese Kokai Tokkyo KohoHl-92659).

[0119] The transdermal absorption parameter was calculated for each testsample. The results are shown in Table 1. In Table 1, AUC [0˜24 hr]represents the area under the blood concentration-time curve over 0˜24hours after application. TABLE 1 Transdermal absorption parameter (n =3, mean ± S.E.) AUC [0-24 hr] Test sample (ng · hr/ml) Formulation 2 >30Formulation 3 >30 Formulation 7 >30 Formulation 12 >30 Formulation 20>30

Effects of the Invention

[0120] In accordance with this invention there can be provided apharmaceutical composition of tricyclic compound (I), particularly ahydrophilic semi-solid composition for external application, which isstable, easy to use, acceptable in feeling in use, and with a lowirritation potential and/or improved dermal penetration.

[0121] The pharmaceutical composition of the present invention is usefulfor the treatment or prevention of Inflammatory or hyperproliferativeskin diseases or cutaneous manifestations of immunologically-mediateddiseases (e.g. psoriasis, atopic dermatitis, contact dermatitis,eczematoid dermatitis, seborrheic dermatitis, lichen planus, pemphigus,bullous pemphigoid, epidermolysis bullosa, urticaria, angioedema,vasculitides, erythema, dermal eosinophilia, lupus erythematosus, acne,and alopecia areata) because of the pharmacologic activities possessedby the tricyclic compound (I).

[0122] Furthermore, the active ingredient, tricyclic compound (I), usedin the pharmaceutical composition is useful for the therapy orprophylaxis of the following diseases.

[0123] Rejection reactions by transplantation of organs or tissues suchas the heart, kidney, liver, bone marrow, skins cornea, lung, pancreas,small intestine, limb, muscle, nerve, intervertebral disc, trachea,myoblast, cartilage, etc.;

[0124] graft-versus-host reactions following bone marrowtransplantation;

[0125] autoimmune diseases such as rheumatoid arthritis, systemic lupuserythematosus, Hashimoto's thyroiditis, multiple sclerosis, myastheniagravis, type I diabetes, etc.;

[0126] and infections caused by pathogenic microorganisms (e.g.Aspergillus fumigatus, Fusarium oxysporum, Trichophyton asteroides,etc.);

[0127] autoimmune diseases of the eye (e.g. keratoconjunctivitis, vernalconjunctivitis, uveitis associated with Behcet's disease, keratitis,herpetic keratitis, conical keratitis, corneal epithelial dystrophy,keratoleukoma, ocular premphigus, Mooren's ulcer, scleritis, Graves'ophthalmopathy, Vogt-Koyanagi-Harada syndrome, keratoconjunctivitissicca (dry eye), phlyctenule, iridocyclitis, sarcoidosis, endocrineophthalmopathy, etc.);

[0128] reversible obstructive airways diseases [asthma (e.g. bronchialasthma, allergicasthma, intrinsicasthma, extrinsic asthma, and dustasthma), particularly chronic or inveterate asthma (e.g. late asthma andairway hyper-responsiveness) bronchitis, etc.] mucosal or vascularinflammations (e.g. gastric ulcer, ischemic or thrombotic vascularinjury, ischemic bowel diseases, enteritis, necrotizing enterocolitis,intestinal damages associated with thermal burns, leukotrieneB4-mediated diseases);

[0129] intestinal inflammations/allergies (e.g. coeliac diseases,proctitis, eosinophilic gastroenteritis, mastocytosis, Crohn's diseaseand ulcerative colitis);

[0130] food-related allergic diseases with symptomatic manifestationremote from the gastrointestinal tract (e.g. migrain, rhinitis andeczema);

[0131] renal diseases (e.g. intestitial nephritis, Goodpasture'ssyndrome, hemolytic uremic syndrome, and diabetic nephropathy);

[0132] nervous diseases (e.g. multiple myositis, Guillain-Barresyndrome, Meniere's disease, multiple neuritis, solitary neuritis,cerebral infarction, Alzheimer's disease, Parkinson's disease,amyotrophic lateral sclerosis (ALS) and radiculopathy);

[0133] cerebral ischemic disease (e.g., head injury, hemorrhage in brain(e.g., subarachnoid hemorrhage, intracerebral hemorrhage) cerebralthrombosis, cerebral embolism, cardiac arrest, stroke, transientischemic attack (TIA), hypertensive encephalopathy);

[0134] endocrine diseases (e.g. hyperthyroidism, and Basedow's disease);

[0135] hematic diseases (e.g. pure red cell aplasia, aplastic anemia,hypoplastic anemia, idiopathic thrombocytopenic purpura, autoimmunehemolytic anemia, agranulocytosis, pernicious anemia, megaloblasticanemia, and anerythroplasia);

[0136] bone diseases (e.g. osteoporosis);

[0137] respiratory diseases (e.g. sarcoidosis, pulmonary fibrosis, andidiopathic interstitial pneumonia);

[0138] skin diseases (e.g. dermatomyositis, leukoderma vulgaris,ichthyosis vulgaris, photosensitivity, and cutaneous T-cell lymphoma);

[0139] circulatory diseases (e.g. arteriosclerosis, atherosclerosis,aortitis syndrome, polyarteritis nodosa, and myocardosis);

[0140] collagen diseases (e.g. scleroderma, Wegener's granuloma, andSjogren's syndrome);

[0141] adiposis;

[0142] eosinophilic fasciitis;

[0143] periodontal diseases (e.g. damage to gingiva, periodontium,alveolar bone or substantia ossea dentis);

[0144] nephrotic syndrome (e.g. glomerulonephritis);

[0145] male pattern alopecia, alopecia senile;

[0146] muscular dystrophy;

[0147] pyoderma and Sezary syndrome;

[0148] Addison's disease;

[0149] chromosome abnormality-associated diseases (e.g. Down'ssyndrome);

[0150] active oxygen-mediated diseases [e.g. organ injury (e.g. ischemiccirculation disorders of organs (e.g. heart, liver, kidney, digestivetract, etc.) associated with preservation, transplantation, or ischemicdiseases (e.g. thrombosis, cardial infarction, etc.)):

[0151] intestinal diseases (e.g. endotoxin shock, pseudomembranouscolitis, and drug- or radiation-induced colitis):

[0152] renal diseases (e.g. ischemic acute renal insufficiency, chronicrenal failure):

[0153] pulmonary diseases (e.g. toxicosis caused by pulmonary oxygen ordrugs (e.g. paracort, bleomycin, etc.), lung cancer, and pulmonaryemphysema):

[0154] ocular diseases (e.g. cataracta, iron-storage disease (siderosisbulbi), retinitis, pigmentosa, senile plaques, vitreous scarring,corneal alkali burn):

[0155] dermatitis (e.g. erythema multiforme, linear immunoglobulin Abullous dermatitis, cement dermatitis):

[0156] and other diseases (e.g. gingivitis, periodontitis, sepsis,pancreatitis, and diseases caused by environmental pollution (e.g. airpollution) , aging, carcinogen, metastasis of carcinoma, andhypobaropathy)];

[0157] diseases caused by histamine release or leukotriene C4 release;

[0158] restenosis of coronary artery following angioplasty andprevention of postsurgical adhesions;

[0159] Autoimmune diseases and inflammatory conditions (e.g., primarymucosal edema, autoimmune atrophic gastritis, premature menopause, malesterility, juvenile diabetes mellitus, pemphigus vulgaris, pemphigoid,sympathetic ophthalmitis, lens-induced uveitis, idiopathic leukopenia,active chronic hepatitis, idiopathic cirrhosis, discoid lupuserythematosus, autoimmune orchitis, arthritis (e.g. arthritisdeformans), or polychondritis);

[0160] Human Immunodeficiency Virus (HIV) infections AIDS; allergicconjunctivitis;

[0161] hypertrophic cicatrix or keloid due to trauma, burn, or surgery.

[0162] In addition, the tricyclic compound (I) has liver regeneratingactivity and/or activities of stimulating hypertrophy and hyperplasia ofhepatocytes.

[0163] Therefore, the pharmaceutical composition of the presentinvention is useful for the therapy or prophylaxis of liver diseases[e.g. immunogenic diseases (e.g. chronic autoimmune liver diseases suchas autoimmune hepatic diseases, primary biliary cirrhosis or sclerosingcholangitis), partial liver resection, acute liver necrosis (e.g.necrosis caused by toxins, viral hepatitis, shock, or anoxia), hepatitisB, non-A non-B hepatitis, hepatocirrhosis, and hepatic failure (e.g.fulminant hepatitis, late-onset hepatitis and “acute-on-chronic” liverfailure (acute liver failure on chronic liver diseases))].

[0164] And further, the present composition is useful for preventing ortreating various diseases because of its useful pharmacological activitysuch as augmenting activity of chemotherapeutic effect, activity ofcytomegalovirus infection, anti-inflammatory activity, inhibitingactivity against peptidyl-prolyl isomerase or rotamase, antimalarialactivity, antitumor activity, and so on.

[0165] Among various formulations for the pharmaceutical composition ofthis invention, those formulations of reduced dermal irritancy are ofvalue for treating or preventing atopic and other diseases of the skin,while those formulations with high dermal absorbability are particularlyuseful for treating or preventing psoriasis and other diseases of theskin.

1. A pharmaceutical composition, which comprises a tricyclic compound ofthe formula:

(wherein each of adjacent pairs of R¹ and R², R³ and R⁴ or R⁵ and R⁶independently (a) is two adjacent hydrogen atoms, or (b) may formanother bond formed between the carbon atoms to which they are attached,and further, R² may be an alkyl group; R⁷ is a hydrogen atom, a hydroxygroup, a protected hydroxy group or an alkoxy group, or an oxo grouptogether with R¹; each of R⁸ and R⁹ is independently a hydrogen atom ora hydroxy group; R¹⁰ is a hydrogen atom, an alkyl group, an alkyl groupsubstituted by one or more hydroxy groups, an alkenyl group, an alkenylgroup substituted by one or more hydroxy groups, or an alkyl groupsubstituted by an oxo group; X is an oxo group, (a hydrogen atom and ahydroxy group), (a hydrogen atom and a hydrogen atom), or a grouprepresented by the formula —CH₂O—; Y is an oxo group, (a hydrogen atomand a hydroxy group), (a hydrogen atom and a hydrogen atom), or a grouprepresented by the formula N—NR¹¹R¹² or N—OR¹³; each of R¹¹ and R¹² isindependently a hydrogen atom, an alkyl group, an aryl group or a tosylgroup; each of R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, R²² and R²³ isindependently a hydrogen atom or an alkyl group; each of R²⁰ and R²¹ isindependently an oxo group or (R²⁰a and a hydrogen atom) or (R²¹a and ahydrogen atom) in which each of R²⁰a and R²¹a is independently a hydroxygroup, an alkoxy group or a group represented by the formula—OCH₂OCH₂CH₂OCH₃, or R²¹a is a protected hydroxy group, or R²⁰a and R²¹amay together represent an oxygen atom in an epoxide ring; n is aninteger of 1 or 2; and in addition to the above definitions, Y, R¹⁰ andR²³, together with the carbon atoms to which they are attached, mayrepresent a saturated or unsaturated 5- or 6-membered nitrogen, sulfurand/or oxygen containing heterocyclic ring optionally substituted by oneor more groups selected from the group consisting of an alkyl, ahydroxy, an alkyl substituted by one or more hydroxy groups, an alkoxy,a benzyl and a group of the formula —CH₂Se (C₆H₅)), or itspharmaceutically acceptable salt, an oil substance, a surfactant, ahydrophilic substance, and water, and further optionally a pH controlagent.
 2. The pharmaceutical composition according to claim 1, whereinthe surfactant is polyoxyethylene alkyl ether, polyoxyethylene sorbitanfatty acid ester, polyethylene glycol fatty acid ester, pentaglycerolfatty acid ester, or glycerol fatty acid ester.
 3. The pharmaceuticalcomposition according to claim 2, wherein the amount of the surfactant,the oil substance and the hydrophilic substance are 0.1˜5% (w/w), 2˜50%(w/w), and 0.1˜10% (w/w) among the total composition, respectively. 4.The pharmaceutical composition according to claim 3, wherein the oilsubstance is a dibasic acid diester and/or a monohydric alcohol fattyacid ester.
 5. The pharmaceutical composition according to claim 4,wherein the oil substance is isopropyl myristate and/or diethylsebacate.
 6. The pharmaceutical composition according to claim 5,wherein the hydrophilic substance is a carboxyvinyl polymer.
 7. Thepharmaceutical composition according to claim 1, wherein the tricycliccompound (I) is the one therein each of adjacent pairs of R³ and R⁴ orR⁵ and R⁶ independently may form another bond formed between the carbonatoms to which they are attached; each of R⁸ and R²³ is independently ahydrogen atom; R⁹ is a hydroxy group; R¹⁰ is a methyl group, an ethylgroup, a propyl group or an allyl group; X is (a hydrogen atom and ahydrogen atom) or an oxo group; Y is an oxo group; each of R¹⁴, R¹⁵,R¹⁶, R¹⁷, R¹⁸, R¹⁹, and R²² is a methyl group; each of R²⁰ and R²¹ isindependently (R²⁰a and a hydrogen atom) or (R²¹a and a hydrogen atom)in which each of R²⁰a and R²¹a is a hydroxy group or an alkoxy group, orR²¹a is a protected hydroxy group; and n is an integer of 1 or
 2. 8. Thepharmaceutical composition according to claim 7, wherein said tricycliccompound (I) is17-allyl-1,14-dihydroxy-12-[2-(4-hydroxy-3-methoxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl-11,28-dioxa-4-azatricyclo[22.3.1.0^(4,9)]octacos-18-ene-2,3,10,16-tetraone.9. The pharmaceutical composition according to claim 8, which has beenadjusted to pH about 4˜5.
 10. A process for producing the pharmaceuticalcomposition claimed in claim 1, which comprises mixing a solutioncomprising a tricyclic compound or a pharmaceutically acceptable saltthereof, an oil substance and a surfactant, with water to prepare anemulsion; and then mixing it with a hydrophilic substance and furtheroptionally a pH control agent.
 11. The process according to claim 10,which comprises mixing the hydrophilic substance and optionally a pHcontrol agent, with water separately; and then mixing it with theemulsion defined in claim 10.